| Solution Information | help | |
| Enzyme: | Phosphoethanolamine/phosphocholine phosphatase | |
| inhibitor: | BDBM46043 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Assay materials: 1) POSPHO1 was obtained from the assay provider's laboratory. 2) Assay Buffer: 20mM MES-NaOH pH 6.7, 2mM MgCl2, 0.0125% Tween-20, 0.01% BSA Procedure 1) Dose-response curves contained 10 concentrations of compounds obtained using 2-fold serial dilution. Compounds were serially diluted in 100% DMSO, and then diluted with water to 10% final DMSO concentration. 2 uL of compounds in 10% DMSO were transferred into columns 3-22 of Greiner 384-well white small-volume plates (784075). Each curve was performed in duplicate. 2) Columns 1-2 and 23-24 contained 4 uL of NC and PC solutions, respectively. For the following additions, using the Multidrop sterilizing the tubing with copious amounts of water and 70% EtOH before and after each reagent. 3) Add 9 uL of buffer to columns 1-2. 4) Add 9 uL PHOSPHO1 intermediate solution to columns 3-24. 5) Add 9 uL of PEA intermediate solution to the entire plate (columns 1-24). 6) Spin plate down to maintain an even level of volume | |
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